Application of hyper-branched rolling circle amplification (HRCA) and HRCA-based strip test for the detection of Chattonella marina

Environmental Science and Pollution Research International
Xiaoli NieGuofu Chen

Abstract

Harmful algal blooms (HABs) are global threats to marine ecosystems, fisheries, and human health. Therefore, developing effective and accurate methods for identifying causative algae and monitoring seawater quality is urgent. However, traditional, microscopy-based methods are complex, inaccurate, and time-consuming. Here, we present a novel method for effective and sensitive detection of Chattonella marina using hyper-branched rolling circle amplification (HRCA) and HRCA-based strip test (HBST). The large subunit (LSU) ribosomal DNA (rDNA) D1-D2 region of C. marina was firstly sequenced to design a species-specific padlock probe (PLP). The HRCA reaction with two amplification primers and further HBST for C. marina was established. The optimized reaction conditions for HRCA were PLP concentration, 20 pM; ligation temperature, 65 °C; ligation time, 60 min; amplification temperature, 61 °C; and amplification time, 60 min. The developed HBST detection procedure involved HRCA reaction, test strip preparation, hybridization, coloration, and judgment of hybridization by the naked eye. Specificity and sensitivity of the established methods were validated. Moreover, the results showed that the established detection methods were specific...Continue Reading

References

Apr 21, 2001·BioTechniques·G J HafnerP M Giffard
Mar 31, 2012·Annual Review of Marine Science·Donald M AndersonGustaaf M Hallegraeff
Dec 18, 2013·Environmental Science and Pollution Research International·Chunyun ZhangGuangce Wang
Jun 1, 2006·Oceanography : the Official Magazine of the Oceanography Society·Lorraine C Backer, Dennis J McGillicuddy

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Citations

Aug 11, 2020·Environmental Science and Pollution Research International·Fuguo LiuGuofu Chen
Aug 23, 2020·The Science of the Total Environment·Anna ToldràMònica Campàs
Apr 5, 2021·Environmental Science and Pollution Research International·Fuguo LiuGuofu Chen

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